As is known, the analysis of nucleic acids requires, according to different modalities, preliminary steps of preparation of a sample of biological material, of amplification of the nucleic material contained therein, and of hybridization of individual target or reference strands, corresponding to the sequences sought. Hybridization occurs (and the test yields a positive outcome) if the sample contains strands complementary to the target strands.
At the end of the preparatory steps, the sample must be examined to control whether hybridization has occurred (the so called detection step). For this purpose, various inspection methods and apparatuses are known, for example of an optical or electrical type. In particular, the methods and apparatuses of an optical type are frequently based upon the phenomenon of fluorescence. The reactions of amplification and hybridization are conducted so that the hybridized strands, contained in a detection chamber made in a support, include fluorescent molecules or fluorofors (the hybridized strands may be either grafted to the bottom of the detection chamber or remain in liquid suspension). The support is exposed to a light source having an appropriate spectrum of emission, such as to excite the fluorofors. In turn, the excited fluorofors emit a secondary radiation at an emission wavelength higher than the peak of the excitation spectrum. The light emitted by the fluorofors is collected and captured by an optical sensor. In order to eliminate the background light radiation, which represents a source of disturbance, the optical sensor is provided with band-pass or interferential filters centred at the wavelength of emission of the fluorofors.
However, the difference between the maximum peak of the emission spectrum of the fluorofors and the peak of the excitation spectrum (also referred to as “Stokes shift”) is not very high, and the filters, however selective they may be, can only attenuate the light emitted by the source and subsequently diffused, without, however, eliminating it altogether. It should also be taken into account that the materials used for providing the supports often have high reflecting power. For example, microfluidic devices for the analysis of nucleic acids integrated in semiconductor chips are increasingly widespread. In integrated microfluidic devices, the detection chamber often has the bottom coated with a layer of silicon dioxide and, sometimes, also metal electrodes are present, for example of gold or aluminium. In effect, hence, only a relatively small part of the light emitted by the source is absorbed, whereas a conspicuous fraction is reflected and is potentially capable of disturbing the detection of the light emitted by the fluorofors.